Compositions and Methods for Dengue Virus Chimeric Constructions in Vaccines

A dengue-1/dengue-2 chimera, wherein the dengue-1/dengue-2 chimera

• is encoded by a polynucleotide molecule encoding a dengue-1/dengue-2 polypeptide chimera,
• wherein the polynucleotide molecule encoding the dengue-1/dengue-2 polypeptide chimera comprises a first nucleotide sequence encoding nonstructural proteins from a modified live, attenuated dengue-2 virus strain PDK 53, a second nucleotide sequence encoding at least one structural protein from dengue-1, and at least one mutation, wherein the at least one mutation comprises one or more of:
• an adenine to cytosine mutation at position 3823 in the numbering of SEQ ID NO:
• 13 encoding a leucine instead of an isoleucine in the dengue-1/dengue-2 polypeptide chimera at amino acid position 1243 in the numbering of SEQ ID NO:
• 3 corresponding to NS2A-116;
• an adenine to thymine mutation at position 4407 in the numbering of SEQ ID NO:
• 13 encoding an aspartic acid instead of a glutamic acid in the dengue-1/dengue-2 polypeptide chimera at amino acid position 1437 in the numbering of SEQ ID NO:
• 3 corresponding to NS2B-92;
• and
• an adenine to guanine mutation at position 7311.

A dengue-1/dengue-2 chimera, wherein the dengue-1/dengue-2 chimera

• comprises an RNA transcribed from a cDNA comprising a polynucleotide molecule encoding the dengue-1/dengue-2 polypeptide chimera,
• wherein the polynucleotide molecule encoding the dengue-1/dengue-2 polypeptide chimera comprises a first nucleotide sequence encoding nonstructural proteins from a modified live, attenuated dengue-2 virus strain PDK 53, a second nucleotide sequence encoding at least one structural protein from dengue-1, and at least one mutation, wherein the at least one mutation comprises one or more of:
• an adenine to cytosine mutation at position 3823 in the numbering of SEQ ID NO:
• 13 encoding a leucine instead of an isoleucine in the dengue-1/dengue-2 polypeptide chimera at amino acid position 1243 in the numbering of SEQ ID NO:
• 3 corresponding to NS2A-116;
• an adenine to thymine mutation at position 4407 in the numbering of SEQ ID NO:
• 13 encoding an aspartic acid instead of a glutamic acid in the dengue-1/dengue-2 polypeptide chimera at amino acid position 1437 in the numbering of SEQ ID NO:
• 3 corresponding to NS2B-92;
• and
• an adenine to guanine mutation at position 7311.

A dengue-1/dengue-2 chimera, wherein the dengue-1/dengue-2 chimera

• comprises one or more polypeptide molecules encoded by a polynucleotide molecule encoding the dengue-1/dengue-2 polypeptide chimera,
• wherein the polynucleotide molecule encoding the dengue-1/dengue-2 polypeptide chimera comprises a first nucleotide sequence encoding nonstructural proteins from a modified live, attenuated dengue-2 virus strain PDK 53, a second nucleotide sequence encoding at least one structural protein from dengue-1, and at least one mutation, wherein the at least one mutation comprises one or more of:
• an adenine to cytosine mutation at position 3823 in the numbering of SEQ ID NO:
• 13 encoding a leucine instead of an isoleucine in the dengue-1/dengue-2 polypeptide chimera at amino acid position 1243 in the numbering of SEQ ID NO:
• 3 corresponding to NS2A-116;
• an adenine to thymine mutation at position 4407 in the numbering of SEQ ID NO:
• 13 encoding an aspartic acid instead of a glutamic acid in the dengue-1/dengue-2 polypeptide chimera at amino acid position 1437 in the numbering of SEQ ID NO:
• 3 corresponding to NS2B-92;
• and an adenine to guanine mutation at position 7311.

A dengue-1/dengue-2 chimera, wherein the dengue-1/dengue-2 chimera

• is obtainable by a method for producing a dengue-1/dengue-2 chimera, the method comprising the following steps:
• a) transcribing an RNA from a cDNA comprising a polynucleotide molecule encoding the dengue-1/dengue-2 polypeptide chimera, and
• b) introducing the RNA transcribed in step a) into cells for production of the dengue-1/dengue-2 chimera,
• wherein the polynucleotide molecule encoding the dengue-1/dengue-2 polypeptide chimera comprises a first nucleotide sequence encoding nonstructural proteins from a modified live, attenuated dengue-2 virus strain PDK 53, a second nucleotide sequence encoding at least one structural protein from dengue-1, and at least one mutation, wherein the at least one mutation comprises one or more of:
• an adenine to cytosine mutation at position 3823 in the numbering of SEQ ID NO:
• 13 encoding a leucine instead of an isoleucine in the dengue-1/dengue-2 polypeptide chimera at amino acid position 1243 in the numbering of SEQ ID NO:
• 3 corresponding to NS2A-116;
• an adenine to thymine mutation at position 4407 in the numbering of SEQ ID NO:
• 13 encoding an aspartic acid instead of a glutamic acid in the dengue-1/dengue-2 polypeptide chimera at amino acid position 1437 in the numbering of SEQ ID NO:
• 3 corresponding to NS2B-92;
• and an adenine to guanine mutation at position 7311.